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Transient Fluorescence Labeling: Low Affinity—High Benefits

Fluorescent labeling is an established method for visualizing cellular structures and dynamics. The fundamental diffraction limit in image resolution was recently bypassed with the development of super-resolution microscopy. Notably, both localization microscopy and stimulated emission depletion (ST...

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Detalles Bibliográficos
Autores principales: Perfilov, Maxim M., Gavrikov, Alexey S., Lukyanov, Konstantin A., Mishin, Alexander S.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2021
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8583718/
https://www.ncbi.nlm.nih.gov/pubmed/34769228
http://dx.doi.org/10.3390/ijms222111799
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author Perfilov, Maxim M.
Gavrikov, Alexey S.
Lukyanov, Konstantin A.
Mishin, Alexander S.
author_facet Perfilov, Maxim M.
Gavrikov, Alexey S.
Lukyanov, Konstantin A.
Mishin, Alexander S.
author_sort Perfilov, Maxim M.
collection PubMed
description Fluorescent labeling is an established method for visualizing cellular structures and dynamics. The fundamental diffraction limit in image resolution was recently bypassed with the development of super-resolution microscopy. Notably, both localization microscopy and stimulated emission depletion (STED) microscopy impose tight restrictions on the physico-chemical properties of labels. One of them—the requirement for high photostability—can be satisfied by transiently interacting labels: a constant supply of transient labels from a medium replenishes the loss in the signal caused by photobleaching. Moreover, exchangeable tags are less likely to hinder the intrinsic dynamics and cellular functions of labeled molecules. Low-affinity labels may be used both for fixed and living cells in a range of nanoscopy modalities. Nevertheless, the design of optimal labeling and imaging protocols with these novel tags remains tricky. In this review, we highlight the pros and cons of a wide variety of transiently interacting labels. We further discuss the state of the art and future perspectives of low-affinity labeling methods.
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spelling pubmed-85837182021-11-12 Transient Fluorescence Labeling: Low Affinity—High Benefits Perfilov, Maxim M. Gavrikov, Alexey S. Lukyanov, Konstantin A. Mishin, Alexander S. Int J Mol Sci Review Fluorescent labeling is an established method for visualizing cellular structures and dynamics. The fundamental diffraction limit in image resolution was recently bypassed with the development of super-resolution microscopy. Notably, both localization microscopy and stimulated emission depletion (STED) microscopy impose tight restrictions on the physico-chemical properties of labels. One of them—the requirement for high photostability—can be satisfied by transiently interacting labels: a constant supply of transient labels from a medium replenishes the loss in the signal caused by photobleaching. Moreover, exchangeable tags are less likely to hinder the intrinsic dynamics and cellular functions of labeled molecules. Low-affinity labels may be used both for fixed and living cells in a range of nanoscopy modalities. Nevertheless, the design of optimal labeling and imaging protocols with these novel tags remains tricky. In this review, we highlight the pros and cons of a wide variety of transiently interacting labels. We further discuss the state of the art and future perspectives of low-affinity labeling methods. MDPI 2021-10-30 /pmc/articles/PMC8583718/ /pubmed/34769228 http://dx.doi.org/10.3390/ijms222111799 Text en © 2021 by the authors. https://creativecommons.org/licenses/by/4.0/Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https://creativecommons.org/licenses/by/4.0/).
spellingShingle Review
Perfilov, Maxim M.
Gavrikov, Alexey S.
Lukyanov, Konstantin A.
Mishin, Alexander S.
Transient Fluorescence Labeling: Low Affinity—High Benefits
title Transient Fluorescence Labeling: Low Affinity—High Benefits
title_full Transient Fluorescence Labeling: Low Affinity—High Benefits
title_fullStr Transient Fluorescence Labeling: Low Affinity—High Benefits
title_full_unstemmed Transient Fluorescence Labeling: Low Affinity—High Benefits
title_short Transient Fluorescence Labeling: Low Affinity—High Benefits
title_sort transient fluorescence labeling: low affinity—high benefits
topic Review
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8583718/
https://www.ncbi.nlm.nih.gov/pubmed/34769228
http://dx.doi.org/10.3390/ijms222111799
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